WO2021246666A1 - Anticancer drug composition comprising as active ingredient tlr5 agonist derived from flagellin - Google Patents
Anticancer drug composition comprising as active ingredient tlr5 agonist derived from flagellin Download PDFInfo
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- WO2021246666A1 WO2021246666A1 PCT/KR2021/005734 KR2021005734W WO2021246666A1 WO 2021246666 A1 WO2021246666 A1 WO 2021246666A1 KR 2021005734 W KR2021005734 W KR 2021005734W WO 2021246666 A1 WO2021246666 A1 WO 2021246666A1
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- flagellin
- tlr5 agonist
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Definitions
- the present invention relates to an anticancer composition
- an anticancer composition comprising a flagellin-derived TLR5 agonist as an active ingredient.
- the national R&D project that supported the present invention is based on project specific number 1465029638, project number HI14C3417020019, department name Ministry of Health and Welfare, project management agency name Korea Health Industry Promotion Agency, research project name Leading specialized research project (R&D), research project name small molecule free radical removal material New drug development for the treatment of immune and non-immune tissue damage, the research period is 2019-08-01 ⁇ 2019-11-30.
- the national R&D project that supported the present invention is project specific number 1345331440, project number 2020R1I1A1A01067669, department name Ministry of Education, project management organization name National Research Foundation, research project name Science and Engineering Research Foundation (R&D), research project name recurrence/non-response
- R&D research project name recurrence/non-response
- TLR5 Toll-like receptor 5
- PNAS. 95 (2): 588-93 is a protein encoded by the TLR5 gene in humans (PNAS. 95 (2): 588-93), and is a member of the toll-like receptor (TLR) family. It is known that TLR5 can recognize flagellin of invading motile bacteria (Seminars in Immunopathology. 29 (3): 275-88) TLR5 is involved in the initiation of various diseases, including inflammatory bowel disease. known (Journal of Physiology and Pharmacology. 60 Suppl 4: 71-5).
- Flagellin is a major structural protein constituting the filaments of the bacterial flagella, a motor organelle. Tens of thousands of flagellin molecules polymerize spirally to form long whip-shaped flagella filaments. Flagellin includes a D0 domain, a D1 domain, a D2 domain, and a D3 domain, of which the D0 domain and the D1 domain are required for filament assembly. The D0 and D1 domains of flagellin are highly conserved in structure and sequence across a variety of bacterial species and function as a common molecular pattern in flagellar bacteria that informs the host of bacterial infection. Flagellin is recognized by TLR5 and is known to activate the NF-kB signaling mechanism to induce innate immune stimulation, cytoprotection and radioresistance.
- cancer is one of the diseases that occupies the largest proportion in the cause of death of modern people. It is a disease caused by changes in normal cells due to mutations in genes caused by various causes. It refers to malignant tumors among non-compliant tumors. Cancer is characterized by "uncontrolled cell growth", and by this abnormal cell growth, a cell mass called a tumor is formed and penetrates into surrounding tissues, and in severe cases, it metastasizes to other organs in the body. . Cancer is an intractable chronic disease that causes pain to the patient and ultimately death, without being able to cure fundamentally in many cases even when treated with surgery, radiation, and drug therapy.
- Drug treatment of cancer that is, anticancer drugs
- the side effects could be lowered, but there was a limitation in that resistance was generated with a high probability. Therefore, in recent years, interest in immuno-cancer drugs that use the body's immune system to reduce problems due to toxicity and resistance is rapidly increasing.
- an immune checkpoint blockade that binds to PD-L1 on the surface of cancer cells and inhibits the binding of T cells to PD-1 to activate T cells and attack cancer cells has been developed.
- the present inventors have developed a novel peptide agonist of TLR5 (Toll-like receptor 5) derived from flagellin, and experimentally confirmed the possibility of using the TLR5 agonist alone or in combination as an anticancer composition. , the present invention has been completed.
- TLR5 Toll-like receptor 5
- It is an object of the present invention to provide an anticancer composition comprising a TLR5 agonist derived from flagellin and a pharmaceutically acceptable carrier.
- an anticancer composition comprising a therapeutically effective amount of a TLR5 agonist derived from flagellin.
- the TLR5 agonist derived from flagellin may include a D0 domain and a D1 domain of flagellin.
- the flagellin-derived TLR5 agonist may include a linker peptide having the amino acid sequence set forth in SEQ ID NO: 1 inside the D1 domain.
- the flagellin-derived TLR5 agonist may include the amino acid sequence set forth in SEQ ID NO:2.
- the flagellin-derived TLR5 agonist may modulate the tumor microenvironment.
- the flagellin-derived TLR5 agonist may increase M1 (F4/80+CD206-) polarization and decrease M2 (F4/80+CD206+) polarization in the spleen and lymph nodes.
- the composition may have a growth inhibitory activity of cancer cells.
- the anticancer composition comprising a therapeutically effective amount of a flagellin-derived TLR5 agonist may further include an immune checkpoint inhibitor.
- the immune checkpoint inhibitor may be an anti-PD-1 antibody.
- the anti-PD-1 antibody is avelumab, trimelimumab, ipilimumab, nivolumab, pembrolizumab, atezolizumab, It may be at least one selected from the group consisting of duvalumab, lambrolizumab, AMP-224, MEDI4376 and CT-011.
- the anticancer composition comprising a therapeutically effective amount of a TLR5 agonist derived from flagellin may include a pharmaceutically acceptable carrier.
- a pharmaceutical preparation comprising an anticancer composition comprising a therapeutically effective amount of a flagellin-derived TLR5 agonist according to the present invention.
- the present invention relates to an anticancer composition
- an anticancer composition comprising a flagellin-derived TLR5 agonist as an active ingredient. Since the flagellin-derived TLR5 agonist of the present invention can exhibit anti-cancer or anti-cancer adjuvant effects either alone or at the same time as an immune checkpoint inhibitor, it can be developed as an active ingredient for inhibiting the growth of cancer cells.
- TLR5 agonist according to the present invention, which is a polypeptide of the amino acid sequence of SEQ ID NO: 2 in a C57BL/6 (H-2kb) colorectal cancer mouse animal model transplanted with the MC-38 cell line (hereinafter, the TLR5 agonist of SEQ ID NO: 2) Also referred to as "KMRC011") and anti-PD-1 antibody administration is a schematic diagram showing the overall experiment.
- FIG. 2 is a graph showing the experimental results showing the tumor size for each administered material after the date of tumor transplantation according to the single or combined administration of the TLR5 agonist and the anti-PD-1 antibody according to the present invention in a colorectal cancer mouse model.
- FIG. 3 is a photograph showing the anticancer inhibitory effect of the TLR5 agonist and the anti-PD-1 antibody according to the present invention alone or in combination in a colorectal cancer mouse model.
- FIG. 4 is a result of flow cytometry analysis of immune profiling in spleen and lymph node cells following single or combined administration of a TLR5 agonist and an anti-PD-1 antibody according to the present invention in a colorectal cancer mouse model.
- FIG. 5 shows changes in M1 polarization and M2 polarization in spleen and lymph node cells following administration of a TLR5 agonist and an anti-PD-1 antibody according to the present invention alone or in combination in a colorectal cancer mouse model.
- FIG. 6 is a schematic diagram showing the overall experiment according to the administration of the TLR5 agonist and anti-PD-1 antibody according to the present invention, which is a polypeptide having the amino acid sequence of SEQ ID NO: 2, to a B-cell lymphoma mouse model.
- FIG. 8 is a graph of experimental results showing the tumor size for each administered substance after the date of tumor transplantation following administration of the TLR5 agonist and the anti-PD-1 antibody according to the present invention alone or in combination in a B-cell lymphoma mouse model.
- FIG. 9 is a graph of experimental results showing the tumor size for each administered material after the date of tumor transplantation following administration of the TLR5 agonist and the anti-PD-1 antibody according to the present invention alone or in combination in a B-cell lymphoma mouse model.
- the present invention provides an anticancer composition comprising a therapeutically effective amount of a TLR5 agonist derived from flagellin.
- TLR5 agonist derived from flagellin is derived from or modified from the bacterial flagellin protein, and is a protein or polypeptide having an activity to activate signal transduction by TLR5 (Toll-like receptor 5). is meant to include all of the
- flagellin refers to a major protein constituting the filaments of bacterial flagella.
- Flagellin comprises a D0 domain, a D1 domain, a D2 domain, and a D3 domain. Flagellin is recognized by TLR5 and is known to activate the NF-kB signaling mechanism to induce innate immune stimulation, cytoprotection and radioresistance.
- the "TLR5 agonist derived from flagellin" peptide material may include a D0 domain of flagellin and a D1 domain of flagellin.
- the "TLR5 agonist derived from flagellin” may include a D0 domain of flagellin, a D1 domain of flagellin, and a linker peptide.
- the linker peptide may be included in the D1 domain.
- the linker peptide may include the amino acid sequence of SEQ ID NO: 1.
- the "TLR5 agonist derived from flagellin” comprises the amino acid sequence of SEQ ID NO:2.
- the "TLR5 agonist derived from flagellin” may exhibit growth inhibitory activity of cancer cells by regulating the tumor microenvironment.
- controlling the tumor microenvironment means increasing M1 (F4/80+CD206-) polarization and decreasing M2 (F4/80+CD206+) polarization in the spleen and lymph nodes.
- cancer is a physiological condition in mammals that is usually characterized by uncontrolled cell growth, including all new cell growth and proliferation (whether malignant or benign), and all precancerous and cancerous means cells and tissues.
- the cancer is melanoma, skin cancer, lung cancer, liver cancer, stomach cancer, pancreatic cancer, bone cancer, head or neck cancer, uterine cancer, ovarian cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hawkins' disease, Esophageal cancer, small intestine cancer, colorectal cancer, colon cancer, rectal cancer, perianal cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, prostate cancer, chronic or acute leukemia, lymphocyte lymphoma, bladder cancer, kidney or ureter cancer, renal cell It may be selected from the group consisting of carcinoma, renal pelvic carcinoma, central nervous system tumor, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma, but is not limited thereto.
- the flagellin-derived TLR5 agonist of the present invention has a preventive or
- the flagellin-derived TLR5 agonist of the present invention has a prophylactic or therapeutic effect on B-cell lymphoma.
- Said B-cell lymphoma is low-grade/follicular non-Hodgkin's lymphoma (NHL), small lymphocytic (SL) NHL, intermediate/follicular NHL, moderate extensive NHL, high-grade immunoblastic NHL, high-grade lymphoblastic NHL, high-grade small undifferentiated Cellular NHL, bulky disease NHL, and Waldenstrom's macroglobulinemia can be selected from the group consisting of, but is not limited thereto.
- composition of the present invention may be provided in the form of an anticancer composition comprising an immune checkpoint inhibitor in a composition comprising a therapeutically effective amount of a TLR5 agonist derived from flagellin.
- the immune system of the living body has an immune checkpoint system to suppress the hyperimmune response caused by the overproliferation of T-cells, and the immune checkpoint functions to suppress the hyperimmune response caused by the overactivation and/or hyperproliferation of T-cells.
- cancer cells exploit the immune checkpoint to prevent T-cells from attacking themselves, thereby evading attack by the immune system, thereby causing cancer.
- the immune checkpoint inhibitor includes an antibody that targets an immune checkpoint protein, which is a protein involved in immune checkpoint, and can treat diseases such as cancer, and the immune checkpoint inhibitor includes an antibody, a fusion It may be a protein, an aptamer, or an immune checkpoint protein-binding fragment thereof.
- the immune checkpoint inhibitor may be an anti-immune checkpoint protein antibody or an antigen-binding fragment thereof.
- the checkpoint inhibitor is an anti-CTLA4 antibody, derivative or antigen-binding fragment thereof; anti-PD-1 antibody, derivative or antigen-binding fragment thereof; anti-LAG-3 antibody, derivative or antigen-binding fragment thereof; anti-OX40 antibody, derivative or antigen-binding fragment thereof; anti-TIM3 antibody, derivative or antigen-binding fragment thereof; and an anti-PD-1 antibody, a derivative thereof, or an antigen-binding fragment thereof.
- avelumab, trimelimumab, ipilimumab, nivolumab, pembrolizumab, atezolizumab, duvalumab It may be at least one selected from the group consisting of lambrolizumab, AMP-224, MEDI4376 and CT-011, but is not limited thereto.
- composition of the present invention may be provided in the form of an anticancer composition comprising a therapeutically effective amount of a TLR5 agonist derived from flagellin and a pharmaceutically acceptable carrier.
- the term "therapeutically effective amount” means an amount suitable for generating an anticancer effect on carcinoma by administering the active ingredient "TLR5 agonist derived from flagellin" to a target patient, and specifically, the “therapeutically effective amount” means a sufficient amount of an agent or compound to be administered that relieves to some extent one or more symptoms of the disease or condition being treated.
- the preferred dosage of the anticancer composition comprising a therapeutically effective amount of the flagellin-derived TLR5 agonist and a pharmaceutically acceptable carrier can be appropriately adjusted.
- the composition may preferably be 50 to 150 ug/kg based on a daily dose.
- composition of the present invention may include a pharmaceutically acceptable carrier in addition to the active ingredient "TLR5 agonist derived from flagellin", and such carriers are commonly used in formulations, including lactose, dextrose, sucrose, Sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate ate, talc, magnesium stearate, menthol and mineral oil, and the like.
- a pharmaceutically acceptable carrier in addition to the active ingredient "TLR5 agonist derived from flagellin”
- such carriers are commonly used in formulations, including lactose, dextrose, sucrose, Sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvin
- composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components.
- a lubricant e.g., a talc, a kaolin, a kaolin, a kaolin, a kaolin, a kaolin, kaolin, kaolin, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, mannitol, mannitol, mannitol, mannitol, mannitol, mannitol, mannitol, mannitol, mannitol
- a suitable dosage of the composition of the present invention may be variously prescribed depending on factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity of the patient. have.
- composition of the present invention may be administered orally or parenterally, and when administered parenterally, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc.
- concentration of the component may be determined in consideration of the purpose of treatment, the condition of the patient, the required period, etc., and is not limited to a concentration within a specific range.
- composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person of ordinary skill in the art to which the present invention pertains, or It can be prepared by pouring into a multi-dose container.
- the formulation may be in the form of a solution, suspension, or emulsion in oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or stabilizer.
- the present invention provides a pharmaceutical formulation comprising an anticancer composition comprising a therapeutically effective amount of a flagellin-derived TLR5 agonist and a pharmaceutically acceptable carrier. .
- the composition of the present invention may be formulated into a conventional formulation in the pharmaceutical field by itself or by combining with a carrier commonly acceptable in the pharmaceutical field.
- the conventional formulation may be various formulations such as tablets, capsules, solutions, suspensions, etc. for oral administration, injections or suspensions, etc., in order to prevent the drug from being decomposed by gastric acid during oral administration. It can be administered by using an antacid in combination or by formulating a solid preparation for oral administration such as a tablet into a preparation coated with an enteric coating.
- the colorectal cancer cell line, MC-38 was purchased from the Korea Cell Line Bank (Seoul, Korea), containing 1% antibiotics (10 U/mL penicillin and 10 g/mL streptomycin; Gibco), 10% heat-inactivated fetal bovine serum ( MC-38 cells were grown in DMEM medium (Gibco, Carlsbad, CA, USA) containing FBS; Gibco).
- the grown MC-38 cell line (1 ⁇ 10 6 ) was resuspended in sterile physiological saline and implanted subcutaneously into C57BL6 mice at a dose of 200 ⁇ l to introduce a tumor mouse model.
- anti-PD-1 was used as a positive control and physiological saline was used as a negative control.
- the anti-PD-1 and TLR5 agonists were co-administered at the same time to confirm the effect of the co-administration.
- the size of the formed tumor was measured at intervals of 2-3 days from the 11th day after transplantation, and was calculated according to the formula of wide 2 X length X 0.5. On days 10 and 26 after transplantation, mice were euthanized and spleens and lymph nodes were collected.
- the spleen and lymph node tissues collected from the mouse were placed between the rough sides of two opaque slide glasses and the slide glasses were rubbed against each other to separate the tissues into single cell units, and the separated splenocytes were treated with an ammonium-chloride-potassium lysis solution (ACK). Lysing Buffer; Gibco) to lyse the red blood cells. The spleen and lymph node cells were then resuspended in RPMI 1640 medium containing 1% antibiotics (10 U/mL penicillin and 10 g/mL streptomycin; Gibco) and 5% heat-inactivated fetal bovine serum (FBS; Gibco). did
- mice splenocytes or lymph node cells were evaluated by flow cytometry.
- spleen and lymph node cells of mice transplanted with MC-38 cell line were washed with staining buffer and resuspended in staining buffer, followed by anti-mouse CD206 PE (BioLegend, San Diego, CA, USA). ) and anti-mouse F4/80 Alexa Fluor 700 (BioLegend, San Diego, CA, USA) for 30 min at 4 ⁇ C.
- spleen and lymph node cells were washed with staining buffer and resuspended in staining buffer. They were then evaluated using flow cytometry analysis in a FACS_LSR Fortessa (BD Pharmingen, San Diego, CA, USA) using FlowJo software (TreeStar, Ashland, OR, USA).
- A20-Luc-GFP a B cell lymphoma cell line, was purchased from Imanis Life Sciences (New York, USA).
- A20-Luc-GFP is the LV-eGFP-P2A-Neo transgene introduced into the A20 cell line, and expresses luciferase and green fluorescent protein.
- Luciferase is an enzyme that produces light by oxidizing luciferin, a chemical found in the cells of bioluminescent organisms, catalytically by ATP.
- the A20-Luc-GFP cell line was cultured in RPMI medium (Gibco, Carlsbad, CA, USA).
- the grown A20-Luc-GFP cell line (1 ⁇ 10 6 ) was resuspended in sterile physiological saline and implanted subcutaneously in BALB/C mice at a dose of 200 ⁇ l to introduce a tumor mouse model.
- anti-PD-1 was used as a positive control and physiological saline was used as a negative control.
- the anti-PD-1 and TLR5 agonists were co-administered at the same time to confirm the effect of the co-administration. From the 9th day after transplantation, the above drug was administered intraperitoneally three times at 3-day intervals. The size of the formed tumor was measured every 3-4 days from the 12th day after transplantation.
- the A20-Luc-GFP cell line suspended in RPMI medium was dispensed in a 96-well plate at a volume of 200 ⁇ l from 1 ⁇ 10 2 cells to 1x10 5 cells, luciferin was injected at a concentration of 150ug/ml, and bioluminescence was measured to determine the The degree of bioluminescence was confirmed.
- Luciferin was intraperitoneally injected into tumor-implanted mice at 150 mg/kg, and the bioluminescence of the luciferase-expressing tumor was measured to determine the degree of tumor growth in vivo. Bioluminescence was measured using an in vivo fluorescence spectrometer (IVIS Lumina XRMS).
- mice transplanted with the MC-38 cell line intraperitoneally were monitored, and the results are shown in FIG. 3 . It can be seen that it can be observed that the size of the cancer in mice is reduced due to administration of the TLR5 agonist of the present invention when compared to the negative or positive control group.
- TLR5 agonist (“KMRC011") of the present invention can be used as an immune checkpoint inhibitor and can be used as an effective anticancer agent, and can be used as an anticancer adjuvant having a synergistic effect in tumor treatment when administered in combination with an existing immune checkpoint inhibitor. it means.
- the immune profiling of mouse spleen and lymph node cells for each administered substance in a tumor mouse model was evaluated by flow cytometry.
- the present inventors established conditions for differentiation of M1 and M2 macrophages from mouse spleen and lymph node cells.
- M1 macrophages are known to have an anticancer effect by showing tumor aggression
- M2 macrophages are known to grow tumors as tumor-supporting macrophages that are friendly to cancer. Therefore, it was confirmed whether the TLR5 agonist of the present invention had an anticancer effect by regulating the tumor microenvironment of the tumor cell line through the change in the degree of polarization of the M1/M2 macrophages.
- M1 and M2 macrophages from mouse spleen and lymph node cells were analyzed using flow cytometry. The polarization was confirmed by observation, and the results are shown in FIGS. 4 and 5 .
- spleen and lymph node cells treated with TLR5 agonist it was confirmed that the polarization of M1 macrophages (F4/80+ CD206-) was increased, while that of M2 macrophages (F4/80+ CD206+) was decreased.
- the TLR5 agonist can regulate the distribution of immune cell subtypes M1 and M2 macrophages in order to increase the response of the immune checkpoint inhibitor.
- the TLR5 agonist (“KMRC011") of the present invention can create an environment in which the antitumor effect can be optimally exerted by changing the tumor microenvironment either alone or in combination with an existing immune checkpoint inhibitor.
- the degree of bioluminescence according to the number of cells was confirmed by measuring the bioluminescence of different cell numbers from 1 ⁇ 10 2 cells to 1x10 5 cells. Accordingly, it was confirmed that the degree of bioluminescence increased in proportion.
- the tumor growth was compared for each administered substance after inducing a tumor mouse model.
- the size of the tumor formed in mice transplanted with the A20-Luc-GFP cell line was measured by BLI, and the results are shown in FIG. 9 .
- the size of the cancer in mice was due to the combined administration ( ⁇ ) of the TLR5 agonist and anti-PD-1 of the present invention. It can be seen that the smaller ones are observable.
- TLR5 agonist (“KMRC011") of the present invention not only exhibits a single antitumor effect, but can also be used as an anticancer adjuvant having a synergistic effect in tumor treatment when administered in combination with an existing immune checkpoint inhibitor.
- the present invention relates to an anticancer composition
- an anticancer composition comprising a flagellin-derived TLR5 agonist as an active ingredient, wherein the flagellin-derived TLR5 agonist of the present invention exhibits an anticancer or anticancer adjuvant effect either alone or at the same time as an immune checkpoint inhibitor.
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Abstract
The present invention relates to an anticancer drug composition comprising, as an active ingredient, TLR5 agonist derived from flagellin. The TLR5 agonist derived from flagellin of the present invention can exhibit an anticancer or anticancer adjuvant effect alone or concurrently with an immune checkpoint inhibitor, and thus, can be developed as a cancer cell growth inhibitory active ingredient.
Description
본 발명은 플라젤린으로부터 유도된 TLR5 작용제를 유효성분으로 포함하는 항암제 조성물에 관한 것이다.The present invention relates to an anticancer composition comprising a flagellin-derived TLR5 agonist as an active ingredient.
본 발명을 지원한 국가연구개발사업은 과제고유번호 1465029638, 과제번호 HI14C3417020019, 부처명 보건복지부, 과제관리 기관명 한국보건산업진흥원, 연구사업명 선도형특성화연구사업(R&D), 연구과제명 저분자 유리기 제거재 기반 면역 및 비면역 조직손상 치료를 위한 신약개발, 연구기간 2019-08-01~2019-11-30 이다.The national R&D project that supported the present invention is based on project specific number 1465029638, project number HI14C3417020019, department name Ministry of Health and Welfare, project management agency name Korea Health Industry Promotion Agency, research project name Leading specialized research project (R&D), research project name small molecule free radical removal material New drug development for the treatment of immune and non-immune tissue damage, the research period is 2019-08-01 ~ 2019-11-30.
또한, 본 발명을 지원한 국가연구개발사업은 과제고유번호 1345331440, 과제번호 2020R1I1A1A01067669, 부처명 교육부, 과제관리 기관명 한국연구재단, 연구사업명 이공학학술연구기반구축(R&D), 연구과제명 재발성/불응성림프종에서 톨유사수용체 리간드를 활용한 자가조혈모세포이식의 면역재구성 가속화 및 잔존종양 제거율향상 연구, 연구기간 2020-06-01~2023-05-31 이다.In addition, the national R&D project that supported the present invention is project specific number 1345331440, project number 2020R1I1A1A01067669, department name Ministry of Education, project management organization name National Research Foundation, research project name Science and Engineering Research Foundation (R&D), research project name recurrence/non-response A study on the acceleration of immune reconstruction and improvement of the removal rate of residual tumors in autologous hematopoietic stem cell transplantation using toll-like receptor ligands in sexuallymphoma. The study period is 2020-06-01 ~ 2023-05-31.
TLR5(Toll-like receptor 5)는 인간에서 TLR5 유전자에 의해 암호화된 단백질로서(PNAS. 95 (2): 588-93), TLR(toll-like receptor) 패밀리의 한 일원이다. TLR5는 침입하는 운동성 박테리아의 플라젤린(flagellin)을 인식할 수 있는 것으로 알려져 있다(Seminars in Immunopathology. 29 (3): 275-88) TLR5는 염증성 장질환을 포함하여 다양한 질환의 개시에 관여하는 것으로 알려져 있다(Journal of Physiology and Pharmacology. 60 Suppl 4: 71-5).Toll-like receptor 5 (TLR5) is a protein encoded by the TLR5 gene in humans (PNAS. 95 (2): 588-93), and is a member of the toll-like receptor (TLR) family. It is known that TLR5 can recognize flagellin of invading motile bacteria (Seminars in Immunopathology. 29 (3): 275-88) TLR5 is involved in the initiation of various diseases, including inflammatory bowel disease. known (Journal of Physiology and Pharmacology. 60 Suppl 4: 71-5).
플라젤린(Flagellin)은 운동 세포소기관인, 박테리아 편모의 필라멘트를 구성하는 주요 구조 단백질이다. 수 만개의 플라젤린 분자가 나선형으로 중합되어 긴 채찍 모양의 플라젤라 필라멘트를 형성한다. 플라젤린은 D0 도메인(domain), D1 도메인, D2 도메인, D3 도메인을 포함하며, 그 중 D0 도메인과 D1 도메인 필라멘트 조립에 필요하다. 플라젤린의 D0 및 D1 도메인은 다양한 박테리아 종에 걸쳐 구조 및 서열이 고도로 보존되어 있으며 숙주에게 박테리아 감염을 알리는, 편모를 가진 박테리아의 공통 분자 패턴으로서 기능을 한다. 플라젤린은 TLR5에 의해 인식되어 NF-kB 신호 전달 메커니즘을 활성화시켜 선천성 면역 자극, 세포 보호 및 방사선 저항성을 유도하는 것으로 알려져 있다.Flagellin is a major structural protein constituting the filaments of the bacterial flagella, a motor organelle. Tens of thousands of flagellin molecules polymerize spirally to form long whip-shaped flagella filaments. Flagellin includes a D0 domain, a D1 domain, a D2 domain, and a D3 domain, of which the D0 domain and the D1 domain are required for filament assembly. The D0 and D1 domains of flagellin are highly conserved in structure and sequence across a variety of bacterial species and function as a common molecular pattern in flagellar bacteria that informs the host of bacterial infection. Flagellin is recognized by TLR5 and is known to activate the NF-kB signaling mechanism to induce innate immune stimulation, cytoprotection and radioresistance.
한편, 암은 현대인의 사망원인에서 가장 큰 비중을 차지하고 있는 질환 중 하나로서 여러가지 원인에 의하여 발생된 유전자의 돌연변이로 인하여 정상세포가 변화하여 발생된 질병으로서, 정상적인 세포의 분화, 증식, 성장 형태 등을 따르지 않는 종양 중 악성인 것을 지칭한다. 암이란, "제어되지 않은 세포성장"으로 특징지어지며, 이러한 비정상적인 세포 성장에 의해 종양(tumor)이라고 불리는 세포 덩어리가 형성되어 주위의 조직으로 침투되고, 심한 경우에는 신체의 다른 기관으로 전이되기도 한다. 암은 수술, 방사선 및 약물 요법 등으로 치료를 하더라도 많은 경우에 근본적인 치유가 되지 못하고 환자에게 고통을 주며 궁극적으로는 죽음에 이르게 하는 난치성 만성질환이다.On the other hand, cancer is one of the diseases that occupies the largest proportion in the cause of death of modern people. It is a disease caused by changes in normal cells due to mutations in genes caused by various causes. It refers to malignant tumors among non-compliant tumors. Cancer is characterized by "uncontrolled cell growth", and by this abnormal cell growth, a cell mass called a tumor is formed and penetrates into surrounding tissues, and in severe cases, it metastasizes to other organs in the body. . Cancer is an intractable chronic disease that causes pain to the patient and ultimately death, without being able to cure fundamentally in many cases even when treated with surgery, radiation, and drug therapy.
암의 약물 치료, 즉, 항암제는 일반적으로 세포 독성을 가지고 있는 화합물로서 암세포를 공격해 사멸시키는 방식으로 암을 치료하는데, 암세포뿐만 아니라 정상세포에도 손상을 주기 때문에 높은 부작용을 나타낸다. 따라서 부작용을 감소시키기 위하여 표적 항암제들이 개발되었다. 그러나 이러한 표적 항암제들의 경우에는 부작용은 낮출 수 있었으나, 높은 확률로 내성이 생긴다는 한계점을 나타내었다. 따라서, 최근에는 체내의 면역체계를 이용하여 독성 및 내성으로 인한 문제점을 감소시키는 면역 항암제에 대한 관심이 급증하고 있는 추세이다. 이러한 면역 항암제의 일례로서 암세포 표면의 PD-L1에 결합하여 T 세포의 PD-1과의 결합을 억제하여 T 세포를 활성화시키고, 암세포를 공격하도록 만드는 면역관문억제제(Immune checkpoint blockade)가 개발되었다.Drug treatment of cancer, that is, anticancer drugs, are generally cytotoxic compounds that attack and kill cancer cells. Therefore, targeted anticancer drugs have been developed to reduce side effects. However, in the case of these targeted anticancer drugs, the side effects could be lowered, but there was a limitation in that resistance was generated with a high probability. Therefore, in recent years, interest in immuno-cancer drugs that use the body's immune system to reduce problems due to toxicity and resistance is rapidly increasing. As an example of such an immune anticancer agent, an immune checkpoint blockade that binds to PD-L1 on the surface of cancer cells and inhibits the binding of T cells to PD-1 to activate T cells and attack cancer cells has been developed.
최근 PD-1 (programmed death-1), CTLA-4 (cytotoxic T-lymphocyte antigen-)등을 표적으로 하는 면역관문억제제의 개발로 종양환자의 치료반응율이 상당히 개선되었으며, 실제 항 PD-1의 투여는 흑색종과 비소세포폐암에서는 40~45% 종양반응율, 요로상피세포암에서는 13~24%의 종양반응율, 재발성/불응성 호지킨 림프종에서는 87%의 반응율과 17%의 완전 관해를 보이는 성과를 나타내었다. 다만, 아직까지 해당 약이 적용되는 암종과 환자는 제한적이고 1차 반응이 나타나더라도 급속하게 재발이 되는 문제가 나타나는 실정이다.With the recent development of immune checkpoint inhibitors targeting PD-1 (programmed death-1) and CTLA-4 (cytotoxic T-lymphocyte antigen-), the treatment response rate of tumor patients has been significantly improved, and the actual administration of anti-PD-1 showed a 40-45% tumor response rate in melanoma and non-small cell lung cancer, a 13-24% tumor response rate in urothelial cell carcinoma, and an 87% response rate and 17% complete remission in relapsed/refractory Hodgkin's lymphoma. was shown. However, the number of carcinomas and patients to which the drug is applied is limited, and even if a primary reaction occurs, a problem of rapid recurrence appears.
이에, 본 발명자들은 플라젤린으로부터 유도된 TLR5(Toll-like receptor 5)의 신규한 펩타이드 작용제(agonist)를 개발하였고, 상기 TLR5 작용제의 단독 또는 병용적 요법으로 항암제 조성물로의 가능성을 실험적으로 확인하고, 본 발명을 완성하게 되었다.Accordingly, the present inventors have developed a novel peptide agonist of TLR5 (Toll-like receptor 5) derived from flagellin, and experimentally confirmed the possibility of using the TLR5 agonist alone or in combination as an anticancer composition. , the present invention has been completed.
본 발명의 목적은 플라젤린으로부터 유도된 TLR5 작용제 및 약학적으로 허용되는 담체를 포함하는 항암제 조성물을 제공하는 것이다.It is an object of the present invention to provide an anticancer composition comprising a TLR5 agonist derived from flagellin and a pharmaceutically acceptable carrier.
본 발명의 다른 목적 및 기술적 특징은 이하의 발명의 설명, 청구범위 및 도면에 의해 보다 구체적으로 제시된다.Other objects and technical features of the present invention are presented in more detail by the following description of the invention, claims and drawings.
본 발명의 일 구현 예에 따르면, 플라젤린으로부터 유도된 TLR5 작용제의 치료학적 유효량을 포함하는 항암제 조성물에 관한 것이다.According to one embodiment of the present invention, it relates to an anticancer composition comprising a therapeutically effective amount of a TLR5 agonist derived from flagellin.
본 발명에서 상기 플라젤린으로부터 유도된 TLR5 작용제는 플라젤린의 D0 도메인 및 D1 도메인을 포함하는 것일 수 있다.In the present invention, the TLR5 agonist derived from flagellin may include a D0 domain and a D1 domain of flagellin.
본 발명에서 상기 플라젤린으로부터 유도된 TLR5 작용제는 D1 도메인 내부에 서열번호 1에 개시된 아미노산 서열의 링커 펩타이드를 포함하는 것일 수 있다.In the present invention, the flagellin-derived TLR5 agonist may include a linker peptide having the amino acid sequence set forth in SEQ ID NO: 1 inside the D1 domain.
본 발명에서 상기 플라젤린으로부터 유도된 TLR5 작용제는 서열번호 2에 개시된 아미노산 서열을 포함하는 것일 수 있다.In the present invention, the flagellin-derived TLR5 agonist may include the amino acid sequence set forth in SEQ ID NO:2.
본 발명에서 상기 플라젤린으로부터 유도된 TLR5 작용제는 종양 미세환경을 조절하는 것일 수 있다.In the present invention, the flagellin-derived TLR5 agonist may modulate the tumor microenvironment.
본 발명에서 상기 플라젤린으로부터 유도된 TLR5 작용제는 비장과 림프절에서 M1 (F4/80+CD206-) 분극을 증가시키고 M2 (F4/80+CD206+) 분극을 감소시키는 것일 수 있다.In the present invention, the flagellin-derived TLR5 agonist may increase M1 (F4/80+CD206-) polarization and decrease M2 (F4/80+CD206+) polarization in the spleen and lymph nodes.
본 발명에서 상기 조성물은 암세포의 성장 저해 활성을 갖는 것일 수 있다.In the present invention, the composition may have a growth inhibitory activity of cancer cells.
본 발명에서 플라젤린으로부터 유도된 TLR5 작용제의 치료학적 유효량을 포함하는 항암제 조성물은 면역관문억제제를 추가로 포함하는 것일 수 있다.In the present invention, the anticancer composition comprising a therapeutically effective amount of a flagellin-derived TLR5 agonist may further include an immune checkpoint inhibitor.
본 발명에서 상기 면역관문억제제는 항 PD-1 항체인 것일 수 있다.In the present invention, the immune checkpoint inhibitor may be an anti-PD-1 antibody.
본 발명에서 상기 항 PD-1 항체는 아벨루맙(avelumab), 트리멜리무맙(Tremelimumab), 이필리무맙(Ipilimumab), 니볼루맙(Nivolumab), 펨브롤리주맙(Pembrolizumav), 아테졸리주맙(Atezolizumab), 두발루맙(Durvalumab), 람브로리주맙(Lamvrolizumab), AMP-224, MEDI4376 및 CT-011로 이루어진 군으로부터 선택되는 1종 이상인 것일 수 있다.In the present invention, the anti-PD-1 antibody is avelumab, trimelimumab, ipilimumab, nivolumab, pembrolizumab, atezolizumab, It may be at least one selected from the group consisting of duvalumab, lambrolizumab, AMP-224, MEDI4376 and CT-011.
본 발명에서 플라젤린으로부터 유도된 TLR5 작용제의 치료학적 유효량을 포함하는 항암제 조성물은 약제학적으로 허용되는 담체를 포함할 수 있다.In the present invention, the anticancer composition comprising a therapeutically effective amount of a TLR5 agonist derived from flagellin may include a pharmaceutically acceptable carrier.
본 발명의 또 다른 구현 예에 따르면, 본 발명에 따른 플라젤린으로부터 유도된 TLR5 작용제의 치료학적 유효량을 포함하는 항암제 조성물을 포함하는 약학적 제제에 관한 것이다.According to another embodiment of the present invention, it relates to a pharmaceutical preparation comprising an anticancer composition comprising a therapeutically effective amount of a flagellin-derived TLR5 agonist according to the present invention.
본 발명은 플라젤린으로부터 유도된 TLR5 작용제를 유효성분으로 포함하는 항암제 조성물에 관한 것이다. 본 발명의 플라젤린으로부터 유도된 TLR5 작용제는 단독 또는 면역관문억제제와 동시에 항암 또는 항암 보조의 효과를 나타낼 수 있으므로 암세포의 성장 저해 활성성분으로 개발될 수 있다.The present invention relates to an anticancer composition comprising a flagellin-derived TLR5 agonist as an active ingredient. Since the flagellin-derived TLR5 agonist of the present invention can exhibit anti-cancer or anti-cancer adjuvant effects either alone or at the same time as an immune checkpoint inhibitor, it can be developed as an active ingredient for inhibiting the growth of cancer cells.
도 1은 MC-38 세포주가 이식된 C57BL/6(H-2kb) 대장암 마우스 동물 모델에 서열번호 2의 아미노산 서열의 폴리펩타이드인 본 발명에 따른 TLR5 작용제(이하, 서열번호 2의 TLR5 작용제를 "KMRC011"로도 지칭함) 및 항 PD-1 항체 투여에 따른 실험 전반을 나타내는 모식도이다.1 is a TLR5 agonist according to the present invention, which is a polypeptide of the amino acid sequence of SEQ ID NO: 2 in a C57BL/6 (H-2kb) colorectal cancer mouse animal model transplanted with the MC-38 cell line (hereinafter, the TLR5 agonist of SEQ ID NO: 2) Also referred to as "KMRC011") and anti-PD-1 antibody administration is a schematic diagram showing the overall experiment.
도 2는 대장암 마우스 모델에서 본 발명에 따른 TLR5 작용제 및 항 PD-1 항체의 단독 또는 병용 투여에 따른 종양 이식 일자 이후 투여 물질 별 종양 크기를 나타낸 실험 결과 그래프이다.2 is a graph showing the experimental results showing the tumor size for each administered material after the date of tumor transplantation according to the single or combined administration of the TLR5 agonist and the anti-PD-1 antibody according to the present invention in a colorectal cancer mouse model.
도 3은 대장암 마우스 모델에서 본 발명에 따른 TLR5 작용제 및 항 PD-1 항체의 단독 또는 병용 투여에 따른 항암 억제 효과를 보여주는 실험 결과 사진이다.3 is a photograph showing the anticancer inhibitory effect of the TLR5 agonist and the anti-PD-1 antibody according to the present invention alone or in combination in a colorectal cancer mouse model.
도 4은 대장암 마우스 모델에서 본 발명에 따른 TLR5 작용제 및 항 PD-1 항체의 단독 또는 병용 투여에 따른 비장과 림프절 세포에서 유세포분석기(flow cytometry)의 면역 프로파일링을 분석한 결과이다.4 is a result of flow cytometry analysis of immune profiling in spleen and lymph node cells following single or combined administration of a TLR5 agonist and an anti-PD-1 antibody according to the present invention in a colorectal cancer mouse model.
도 5는 대장암 마우스 모델에서 본 발명에 따른 TLR5 작용제 및 항 PD-1 항체의 단독 또는 병용 투여에 따른 비장과 림프절 세포에서 M1 분극 및 M2 분극의 변화를 나타낸 것이다.5 shows changes in M1 polarization and M2 polarization in spleen and lymph node cells following administration of a TLR5 agonist and an anti-PD-1 antibody according to the present invention alone or in combination in a colorectal cancer mouse model.
도 6은 B 세포 림프종 마우스 모델에 서열번호 2의 아미노산 서열의 폴리펩타이드인 본 발명에 따른 TLR5 작용제 및 항 PD-1 항체 투여에 따른 실험 전반을 나타내는 모식도이다.6 is a schematic diagram showing the overall experiment according to the administration of the TLR5 agonist and anti-PD-1 antibody according to the present invention, which is a polypeptide having the amino acid sequence of SEQ ID NO: 2, to a B-cell lymphoma mouse model.
도 7은 각기 다른 A20-Luc-GFP 세포수의 생물학적발광을 측정하여 세포수에 따른 생물학적발광 정도를 확인한 결과를 나타낸 것이다.7 shows the results of confirming the degree of bioluminescence according to the number of cells by measuring the bioluminescence of different A20-Luc-GFP cell numbers.
도 8은 B 세포 림프종 마우스 모델에서 본 발명에 따른 TLR5 작용제 및 항 PD-1 항체의 단독 또는 병용 투여에 따른 종양 이식 일자 이후 투여 물질 별 종양 크기를 나타낸 실험 결과 그래프이다.8 is a graph of experimental results showing the tumor size for each administered substance after the date of tumor transplantation following administration of the TLR5 agonist and the anti-PD-1 antibody according to the present invention alone or in combination in a B-cell lymphoma mouse model.
도 9는 B 세포 림프종 마우스 모델에서 본 발명에 따른 TLR5 작용제 및 항 PD-1 항체의 단독 또는 병용 투여에 따른 종양 이식 일자 이후 투여 물질 별 종양 크기를 나타낸 실험 결과 그래프이다.9 is a graph of experimental results showing the tumor size for each administered material after the date of tumor transplantation following administration of the TLR5 agonist and the anti-PD-1 antibody according to the present invention alone or in combination in a B-cell lymphoma mouse model.
이하, 본 발명의 바람직한 실시형태들을 설명한다. 그러나, 본 발명의 실시형태는 여러 가지 다른 형태로 변형될 수 있으며, 본 발명의 범위가 이하 설명하는 실시 형태로 한정되는 것은 아니다. 또한, 본 발명의 실시형태는 당해 기술분야에서 평균적인 지식을 가진 자에게 본 발명을 더욱 완전하게 설명하기 위해서 제공되는 것이다.Hereinafter, preferred embodiments of the present invention will be described. However, the embodiment of the present invention may be modified in various other forms, and the scope of the present invention is not limited to the embodiments described below. In addition, the embodiments of the present invention are provided in order to more completely explain the present invention to those of ordinary skill in the art.
상기와 같은 목적을 달성하기 위하여, 본 발명은 플라젤린으로부터 유도된 TLR5 작용제(agonist)의 치료학적 유효량을 포함하는 항암제 조성물을 제공한다.In order to achieve the above object, the present invention provides an anticancer composition comprising a therapeutically effective amount of a TLR5 agonist derived from flagellin.
본 발명에서 사용되는 용어 "플라젤린으로부터 유도된 TLR5 작용제"는 박테리아의 플라젤린 단백질로부터 유래되거나 이를 변형시킨 것으로서 TLR5(Toll-like receptor 5)에 의한 신호전달을 활성화시키는 활성을 갖는 단백질 또는 폴리펩타이드를 모두 포함하는 의미이다.As used herein, the term "TLR5 agonist derived from flagellin" is derived from or modified from the bacterial flagellin protein, and is a protein or polypeptide having an activity to activate signal transduction by TLR5 (Toll-like receptor 5). is meant to include all of the
본 발명에서 사용되는 용어 "플라젤린(flagellin)"은 박테리아 편모의 필라멘트를 구성하는 주요 단백질을 가리킨다. 플라젤린은 D0 도메인(domain), D1 도메인, D2 도메인, D3 도메인을 포함하여 이루어진다. 플라젤린은 TLR5에 의해 인식되어 NF-kB 신호 전달 메커니즘을 활성화시켜 선천성 면역 자극, 세포 보호 및 방사선 저항성을 유도하는 것으로 알려져 있다.As used herein, the term “flagellin” refers to a major protein constituting the filaments of bacterial flagella. Flagellin comprises a D0 domain, a D1 domain, a D2 domain, and a D3 domain. Flagellin is recognized by TLR5 and is known to activate the NF-kB signaling mechanism to induce innate immune stimulation, cytoprotection and radioresistance.
본 발명의 일 구현예에 따르면, 상기 "플라젤린으로부터 유도된 TLR5 작용제" 펩타이드 물질로서 플라젤린의 D0 도메인 및 플라젤린의 D1 도메인을 포함할 수 있다.According to one embodiment of the present invention, the "TLR5 agonist derived from flagellin" peptide material may include a D0 domain of flagellin and a D1 domain of flagellin.
본 발명의 다른 구현예에 따르면, 상기 "플라젤린으로부터 유도된 TLR5 작용제"는 플라젤린의 D0 도메인, 플라젤린의 D1 도메인, 및 링커 펩타이드를 포함할 수 있다.According to another embodiment of the present invention, the "TLR5 agonist derived from flagellin" may include a D0 domain of flagellin, a D1 domain of flagellin, and a linker peptide.
본 발명의 다른 구현예에 따르면, 상기 링커 펩타이드는 D1 도메인 내부에 포함될 수 있다.According to another embodiment of the present invention, the linker peptide may be included in the D1 domain.
본 발명의 다른 구현예에 따르면, 상기 링커 펩타이드는 서열번호 1의 아미노산 서열을 포함할 수 있다.According to another embodiment of the present invention, the linker peptide may include the amino acid sequence of SEQ ID NO: 1.
본 발명의 다른 구현예에 따르면, 상기 "플라젤린으로부터 유도된 TLR5 작용제"는 서열번호 2의 아미노산 서열을 포함한다.According to another embodiment of the present invention, the "TLR5 agonist derived from flagellin" comprises the amino acid sequence of SEQ ID NO:2.
본 발명의 다른 구현예에 따르면, 상기 "플라젤린으로부터 유도된 TLR5 작용제"는 종양 미세환경을 조절함으로써 암세포의 성장 저해 활성을 나타낼 수 있다. 구체적으로, 상기 "종양 미세환경을 제어하는 것"는 비장과 림프절에서 M1 (F4/80+CD206-) 분극을 증가시키고 M2 (F4/80+CD206+) 분극을 감소시키는 것을 의미한다.According to another embodiment of the present invention, the "TLR5 agonist derived from flagellin" may exhibit growth inhibitory activity of cancer cells by regulating the tumor microenvironment. Specifically, "controlling the tumor microenvironment" means increasing M1 (F4/80+CD206-) polarization and decreasing M2 (F4/80+CD206+) polarization in the spleen and lymph nodes.
본 발명에서 사용되는 용어 "암"은 통상적으로 제어되지 않은 세포 성장을 특징으로 하는 포유동물에서의 생리학적 병증으로, 모든 신생 세포 성장 및 증식(악성이든 또는 양성이든), 및 모든 전암성 및 암성 세포 및 조직을 의미한다.As used herein, the term "cancer" is a physiological condition in mammals that is usually characterized by uncontrolled cell growth, including all new cell growth and proliferation (whether malignant or benign), and all precancerous and cancerous means cells and tissues.
상기 암은 흑색종, 피부암, 폐암, 간암, 위암, 췌장암, 골암, 두부 또는 경부 암, 자궁암, 난소암, 유방암, 나팔관암종, 자궁내막암종, 자궁경부암종, 질암종, 음문암종, 호킨스씨병, 식도암, 소장암, 대장암, 결장암, 직장암, 항문부근암, 내분비선암, 갑상선암, 부갑상선암, 부신암, 연조직 육종, 전립선암, 만성 또는 급성 백혈병, 림프구 림프종, 방광암, 신장 또는 수뇨관암, 신장세포 암종, 신장골반 암종, 중추신경계 종양, 1차 중추신경계 림프종, 척수 종양, 뇌간신경교종 및 뇌하수체 선종으로 이루어진 군에서 선택될 수 있으나, 이에 제한되는 것은 아니다.본 발명의 일 구현예에 따르면, 본 발명의 플라젤린으로부터 유도된 TLR5 작용제는 대장암에 예방 또는 치료 효과를 나타낸다. 상기 대장암은 대장에 생긴 암세포로 이루어진 악성 종양을 의미한다. The cancer is melanoma, skin cancer, lung cancer, liver cancer, stomach cancer, pancreatic cancer, bone cancer, head or neck cancer, uterine cancer, ovarian cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hawkins' disease, Esophageal cancer, small intestine cancer, colorectal cancer, colon cancer, rectal cancer, perianal cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, prostate cancer, chronic or acute leukemia, lymphocyte lymphoma, bladder cancer, kidney or ureter cancer, renal cell It may be selected from the group consisting of carcinoma, renal pelvic carcinoma, central nervous system tumor, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma, but is not limited thereto. According to one embodiment of the present invention, The flagellin-derived TLR5 agonist of the present invention has a preventive or therapeutic effect on colorectal cancer. The colorectal cancer refers to a malignant tumor composed of cancer cells generated in the colon.
본 발명의 일 구현예에 따르면, 본 발명의 플라젤린으로부터 유도된 TLR5 작용제는 B 세포 림프종에 예방 또는 치료 효과를 나타낸다. 상기 B 세포 림프종은 저급/여포성 비-호지킨 림프종(NHL), 소형 림프구성 (SL) NHL, 중급/여포성 NHL, 중급 광범성 NHL, 고급 면역아세포 NHL, 고급 림프아세포 NHL, 고급 소형 미분열 세포 NHL, 거대(bulky) 질환 NHL 및 월덴스트롬(Waldenstrom) 마크로글로불린혈증으로 이루어진 군에서 선택될 수 있으나, 이에 제한되는 것은 아니다.According to one embodiment of the present invention, the flagellin-derived TLR5 agonist of the present invention has a prophylactic or therapeutic effect on B-cell lymphoma. Said B-cell lymphoma is low-grade/follicular non-Hodgkin's lymphoma (NHL), small lymphocytic (SL) NHL, intermediate/follicular NHL, moderate extensive NHL, high-grade immunoblastic NHL, high-grade lymphoblastic NHL, high-grade small undifferentiated Cellular NHL, bulky disease NHL, and Waldenstrom's macroglobulinemia can be selected from the group consisting of, but is not limited thereto.
한편, 본 발명의 조성물은 플라젤린으로부터 유도된 TLR5 작용제의 치료학적 유효량을 포함하는 조성물에 면역관문억제제를 추가로 포함하는 항암제 조성물의 형태로 제공될 수 있다.Meanwhile, the composition of the present invention may be provided in the form of an anticancer composition comprising an immune checkpoint inhibitor in a composition comprising a therapeutically effective amount of a TLR5 agonist derived from flagellin.
생체의 면역 시스템은 T-세포의 과다증식으로 인한 과다면역 반응을 억제하기 위한 면역검문 체계를 가지고 있으며, 면역관문은 T-세포의 과활성화 및/또는 과다증식에 의한 과잉면역 반응을 억제하는 기능을 수행하나, 암 세포는 상기 면역관문을 악용하여 T-세포가 자신을 공격하지 못하도록 함으로써 면역 시스템에 의한 공격을 회피함으로써 암을 유발한다.The immune system of the living body has an immune checkpoint system to suppress the hyperimmune response caused by the overproliferation of T-cells, and the immune checkpoint functions to suppress the hyperimmune response caused by the overactivation and/or hyperproliferation of T-cells. However, cancer cells exploit the immune checkpoint to prevent T-cells from attacking themselves, thereby evading attack by the immune system, thereby causing cancer.
상기 면역관문억제제는 면역관문(immune checkpoint)에 관여되는 단백질인을 면역관문 단백질(immune checkpoint protein)을 표적으로 하는 항체를 포함하는 것으로서 암 등의 질환을 치료할 수 있으며, 면역관문억제제는 항체, 융합 단백질, 압타머 또는 이의 면역 체크포인트 단백질-결합 단편일 수 일 수 있다.The immune checkpoint inhibitor includes an antibody that targets an immune checkpoint protein, which is a protein involved in immune checkpoint, and can treat diseases such as cancer, and the immune checkpoint inhibitor includes an antibody, a fusion It may be a protein, an aptamer, or an immune checkpoint protein-binding fragment thereof.
상기 면역관문억제제는 항-면역관문단백질 항체 또는 이의 항원-결합 단편일 수 있다. 바람직하게, 면역관문억제제는 항-CTLA4 항체, 이의 유도체 또는 이의 항원-결합 단편; 항 PD-1 항체, 이의 유도체 또는 이의 항원-결합 단편; 항-LAG-3 항체, 이의 유도체 또는 이의 항원-결합 단편; 항-OX40 항체, 이의 유도체 또는 이의 항원-결합 단편; 항-TIM3 항체, 이의 유도체 또는 이의 항원-결합 단편; 및 항 PD-1 항체, 이의 유도체 또는 이의 항원-결합 단편 중에서 선택될 수 있다. 더욱 바람직하게, 아벨루맙(avelumab), 트리멜리무맙(Tremelimumab), 이필리무맙(Ipilimumab), 니볼루맙(Nivolumab), 펨브롤리주맙(Pembrolizumav), 아테졸리주맙(Atezolizumab), 두발루맙(Durvalumab), 람브로리주맙(Lamvrolizumab), AMP-224, MEDI4376 및 CT-011로 이루어진 군으로부터 선택되는 1종 이상일 수 있으나 이에 제한되는 것은 아니다.The immune checkpoint inhibitor may be an anti-immune checkpoint protein antibody or an antigen-binding fragment thereof. Preferably, the checkpoint inhibitor is an anti-CTLA4 antibody, derivative or antigen-binding fragment thereof; anti-PD-1 antibody, derivative or antigen-binding fragment thereof; anti-LAG-3 antibody, derivative or antigen-binding fragment thereof; anti-OX40 antibody, derivative or antigen-binding fragment thereof; anti-TIM3 antibody, derivative or antigen-binding fragment thereof; and an anti-PD-1 antibody, a derivative thereof, or an antigen-binding fragment thereof. More preferably, avelumab, trimelimumab, ipilimumab, nivolumab, pembrolizumab, atezolizumab, duvalumab, It may be at least one selected from the group consisting of lambrolizumab, AMP-224, MEDI4376 and CT-011, but is not limited thereto.
본 발명의 조성물은 플라젤린으로부터 유도된 TLR5 작용제의 치료학적 유효량 및 약학적으로 허용되는 담체를 포함하는 항암제 조성물의 형태로 제공될 수 있다.The composition of the present invention may be provided in the form of an anticancer composition comprising a therapeutically effective amount of a TLR5 agonist derived from flagellin and a pharmaceutically acceptable carrier.
본 발명에서 용어 "치료학적 유효량"은 유효성분인 "플라젤린으로부터 유도된 TLR5 작용제"를 대상 환자에게 투여하여 암종에 대한 항암 효과를 일으키기에 적합한 양을 의미하고, 구체적으로 상기 "치료학적 유효량"은 치료되는 질환 또는 상태의 하나 이상의 증상을 어느 정도 완화시키는, 투여되는 약제 또는 화합물의 충분한 양을 의미한다.In the present invention, the term "therapeutically effective amount" means an amount suitable for generating an anticancer effect on carcinoma by administering the active ingredient "TLR5 agonist derived from flagellin" to a target patient, and specifically, the "therapeutically effective amount" means a sufficient amount of an agent or compound to be administered that relieves to some extent one or more symptoms of the disease or condition being treated.
상기 플라젤린으로부터 유도된 TLR5 작용제의 치료학적 유효량 및 약학적으로 허용되는 담체를 포함하는 항암제 조성물의 바람직한 투여량은 적절히 조절될 수 있다. 상기 조성물은 바람직하게 1일 투여량을 기준으로 50 내지 150 ug/kg일 수 있다.The preferred dosage of the anticancer composition comprising a therapeutically effective amount of the flagellin-derived TLR5 agonist and a pharmaceutically acceptable carrier can be appropriately adjusted. The composition may preferably be 50 to 150 ug/kg based on a daily dose.
본 발명의 조성물은 유효성분인 "플라젤린으로부터 유도된 TLR5 작용제" 이외에 약학적으로 허용되는 담체를 포함할 수 있으며, 이러한 담체는 제제시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘, 멘톨 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다.The composition of the present invention may include a pharmaceutically acceptable carrier in addition to the active ingredient "TLR5 agonist derived from flagellin", and such carriers are commonly used in formulations, including lactose, dextrose, sucrose, Sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate ate, talc, magnesium stearate, menthol and mineral oil, and the like.
본 발명의 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다. 적합한 약학적으로 허용되는 담체 및 제제는 Remington's Pharmaceutical Sciences (19th ed., 1995)에 상세히 기재되어 있다.The composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).
본 발명의 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다.A suitable dosage of the composition of the present invention may be variously prescribed depending on factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity of the patient. have.
본 발명의 조성물은 경구 또는 비경구로 투여될 수 있고, 비경구로 투여되는 경우, 정맥내 주입, 피하 주입, 근육 주입, 복강 주입, 경피 투여 등으로 투여될 수 있으며, 본 발명의 조성물에 포함되는 유효성분의 농도는 치료 목적, 환자의 상태, 필요기간 등을 고려하여 결정할 수 있으며 특정 범위의 농도로 한정되지 않는다.The composition of the present invention may be administered orally or parenterally, and when administered parenterally, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc. The concentration of the component may be determined in consideration of the purpose of treatment, the condition of the patient, the required period, etc., and is not limited to a concentration within a specific range.
본 발명의 조성물은 당해 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화 함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기내에 내입시켜 제조될 수 있다. 이때 제형은 오일 또는 수성 매질중의 용액, 현탁액 또는 유화액 형태이거나 엑스제, 분말제, 과립제, 정제 또는 캅셀제 형태일 수도 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person of ordinary skill in the art to which the present invention pertains, or It can be prepared by pouring into a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or stabilizer.
본 발명의 또 다른 목적을 달성하기 위하여, 본 발명은 플라젤린으로부터 유도된 TLR5 작용제(agonist)의 치료학적 유효량 및 약학적으로 허용되는 담체를 포함하는, 항암제 조성물을 포함하는 약학적 제제를 제공한다. In order to achieve another object of the present invention, the present invention provides a pharmaceutical formulation comprising an anticancer composition comprising a therapeutically effective amount of a flagellin-derived TLR5 agonist and a pharmaceutically acceptable carrier. .
상기 목적을 달성하기 위하여 본 발명의 조성물은 그 자체로 또는 약제학적 분야에서 통상적으로 허용되는 담체와 함께 배합하여 약제학적 분야에서 통상적인 제제로 제형화 할 수 있다. 바람직하게, 상기 통상적인 제제는 정제, 캅셀제, 액제, 현탁제 등의 경구투여용제제, 주사용제제 또는 현탁액 등의 다양한 제제일 수 있으며, 경구투여시 약제가 위산에 의해 분해하는 것을 방지하기 위하여 제산제를 병용하거나 정제등의 경구투여용 고형제제를 장용피로 피복한 제제로 제형화하여 투여할 수 있다.In order to achieve the above object, the composition of the present invention may be formulated into a conventional formulation in the pharmaceutical field by itself or by combining with a carrier commonly acceptable in the pharmaceutical field. Preferably, the conventional formulation may be various formulations such as tablets, capsules, solutions, suspensions, etc. for oral administration, injections or suspensions, etc., in order to prevent the drug from being decomposed by gastric acid during oral administration. It can be administered by using an antacid in combination or by formulating a solid preparation for oral administration such as a tablet into a preparation coated with an enteric coating.
실시예 Example
실험 방법experimental method
1. 대장암 마우스 모델 1. Colorectal Cancer Mouse Model
(1) 종양 마우스 모델 확립 및 종양 크기 분석(1) tumor mouse model establishment and tumor size analysis
대장암 세포주인 MC-38를 한국 세포주 은행(한국, 서울)에서 구입하여, 1% 항생제 (10 U/mL 페니실린 및 10 g/mL 스트렙토마이신; Gibco), 10% 열-불활성화 우태아 혈청(FBS; Gibco)을 함유하는 DMEM 배지 (Gibco, Carlsbad, CA, USA)에서 MC-38 세포를 성장시켰다.The colorectal cancer cell line, MC-38, was purchased from the Korea Cell Line Bank (Seoul, Korea), containing 1% antibiotics (10 U/mL penicillin and 10 g/mL streptomycin; Gibco), 10% heat-inactivated fetal bovine serum ( MC-38 cells were grown in DMEM medium (Gibco, Carlsbad, CA, USA) containing FBS; Gibco).
성장된 MC-38 세포주(1 × 10
6)를 멸균생리식염수에 재현탁하여 200μl 용량으로 C57BL6 마우스의 피하에 이식하여 종양 마우스 모델을 도입하였다. 본원발명인 TLR5 작용제의 항암효과를 비교하기 위하여 항 PD-1를 양성 대조군으로, 생리식염수를 음성대조군으로 사용하였다. 또한, 항 PD-1 및 TLR5 작용제를 동시에 병용 투여하여 병용 투여에 따른 효과도 확인하였다. 형성된 종양의 크기는 이식 후 11일째부터 2-3일 간격으로 측정하였고, wide
2 X length X 0.5의 공식에 따라 계산되었다. 이식 후 10일 및 26일째에 마우스를 안락사하여 비장, 림프절을 채취하였다.The grown MC-38 cell line (1 × 10 6 ) was resuspended in sterile physiological saline and implanted subcutaneously into C57BL6 mice at a dose of 200 μl to introduce a tumor mouse model. In order to compare the anticancer effect of the TLR5 agonist of the present invention, anti-PD-1 was used as a positive control and physiological saline was used as a negative control. In addition, the anti-PD-1 and TLR5 agonists were co-administered at the same time to confirm the effect of the co-administration. The size of the formed tumor was measured at intervals of 2-3 days from the 11th day after transplantation, and was calculated according to the formula of wide 2 X length X 0.5. On days 10 and 26 after transplantation, mice were euthanized and spleens and lymph nodes were collected.
(2) 조직 단일세포 분리(2) Tissue single cell isolation
마우스로부터 채취한 비장 및 림프절 조직을 2개의 불투명 슬라이드 글라스의 거친 면 사이에 위치시키고 슬라이드 글라스를 서로 비벼주어 조직을 단일 세포 단위로 분리하고, 분리된 비장세포를 암모늄-클로라이드-포타슘 용해 용액 (ACK Lysing Buffer; Gibco)으로 처리하여 적혈구를 용해하였다. 이후, 비장 및 림프절 세포를 1% 항생제 (10 U/mL 페니실린 및 10 g/mL 스트렙토마이신; Gibco), 5% 열-불활성화 우태아 혈청(FBS; Gibco)을 함유하는 RPMI 1640 배지에 재현탁하였다.The spleen and lymph node tissues collected from the mouse were placed between the rough sides of two opaque slide glasses and the slide glasses were rubbed against each other to separate the tissues into single cell units, and the separated splenocytes were treated with an ammonium-chloride-potassium lysis solution (ACK). Lysing Buffer; Gibco) to lyse the red blood cells. The spleen and lymph node cells were then resuspended in RPMI 1640 medium containing 1% antibiotics (10 U/mL penicillin and 10 g/mL streptomycin; Gibco) and 5% heat-inactivated fetal bovine serum (FBS; Gibco). did
(3) 유세포 분석(3) flow cytometry
마우스의 비장 세포 또는 림프절 세포를 유세포분석기를 통해 평가하였다. 대식세포의 M1 또는 M2 분극을 조사하기 위해 MC-38 세포주를 이식한 마우스의 비장 및 림프절 세포를 염색 완충액으로 세척하고 염색 완충액에 재현탁한 뒤 항-마우스 CD206 PE (BioLegend, San Diego, CA, USA)와 항-마우스 F4/80 Alexa Fluor 700 (BioLegend, San Diego, CA, USA)로 4˚C 에서 30 분간 면역 염색하였다. 염색 후, 비장 및 림프절 세포를 염색 완충액으로 세척하고 염색 완충액에 재현탁하였다. 이후, FlowJo 소프트웨어 (TreeStar, Ashland, OR, USA)를 사용하여 FACS_LSR Fortessa (BD Pharmingen, San Diego, CA, USA)에서 유세포 계측 분석을 사용하여 평가하였다.Mice splenocytes or lymph node cells were evaluated by flow cytometry. To investigate M1 or M2 polarization of macrophages, spleen and lymph node cells of mice transplanted with MC-38 cell line were washed with staining buffer and resuspended in staining buffer, followed by anti-mouse CD206 PE (BioLegend, San Diego, CA, USA). ) and anti-mouse F4/80 Alexa Fluor 700 (BioLegend, San Diego, CA, USA) for 30 min at 4˚C. After staining, spleen and lymph node cells were washed with staining buffer and resuspended in staining buffer. They were then evaluated using flow cytometry analysis in a FACS_LSR Fortessa (BD Pharmingen, San Diego, CA, USA) using FlowJo software (TreeStar, Ashland, OR, USA).
2. B 세포 림프종 마우스 모델2. B-Cell Lymphoma Mouse Model
(1) 종양 세포주(1) tumor cell lines
B 세포 림프종 세포주인 A20-Luc-GFP는 Imanis Life Sciences (미국, 뉴욕)에서 구입하였다. A20-Luc-GFP는 A20 세포주에 LV-eGFP-P2A-Neo 형질전환 유전자를 도입한 것으로, 루시퍼라제(luciferase)와 녹색 형광 단백질을 발현한다. 루시퍼라제는 생물 발광 유기체의 세포에서 발견되는 화학 물질인 루시페린을 ATP의 촉매 효과로 산화시키면서 빛을 생성하는 효소이다. A20-Luc-GFP, a B cell lymphoma cell line, was purchased from Imanis Life Sciences (New York, USA). A20-Luc-GFP is the LV-eGFP-P2A-Neo transgene introduced into the A20 cell line, and expresses luciferase and green fluorescent protein. Luciferase is an enzyme that produces light by oxidizing luciferin, a chemical found in the cells of bioluminescent organisms, catalytically by ATP.
상기 A20-Luc-GFP 세포주를 1% 항생제 (10 U/mL 페니실린 및 10 g/mL 스트렙토마이신; Gibco), 10% 열-불활성화 우태아 혈청(FBS; Gibco)을 함유하는 RPMI 배지 (Gibco, Carlsbad, CA, USA)에서 성장시켰다.The A20-Luc-GFP cell line was cultured in RPMI medium (Gibco, Carlsbad, CA, USA).
(2) 종양 마우스 모델 확립 및 종양 크기 분석(2) tumor mouse model establishment and tumor size analysis
성장된 A20-Luc-GFP 세포주(1Х10
6)를 멸균생리식염수에 재현탁하여 200μl 용량으로 BALB/C 마우스의 피하에 이식하여 종양 마우스 모델을 도입하였다. 본원발명인 TLR5 작용제의 항암효과를 비교하기 위하여 항 PD-1를 양성 대조군으로, 생리식염수를 음성대조군으로 사용하였다. 또한, 항 PD-1 및 TLR5 작용제를 동시에 병용 투여하여 병용 투여에 따른 효과도 확인하였다. 이식한 후 9일째부터 3일 간격으로 총 3회 복강내에 위 약물을 투여하였다. 형성된 종양의 크기는 이식 후 12일째부터 3-4일 간격으로 측정하였다.The grown A20-Luc-GFP cell line (1Х10 6 ) was resuspended in sterile physiological saline and implanted subcutaneously in BALB/C mice at a dose of 200 μl to introduce a tumor mouse model. In order to compare the anticancer effect of the TLR5 agonist of the present invention, anti-PD-1 was used as a positive control and physiological saline was used as a negative control. In addition, the anti-PD-1 and TLR5 agonists were co-administered at the same time to confirm the effect of the co-administration. From the 9th day after transplantation, the above drug was administered intraperitoneally three times at 3-day intervals. The size of the formed tumor was measured every 3-4 days from the 12th day after transplantation.
(3) 생물학적발광 분석(3) bioluminescence analysis
RPMI 배지에 현탁시킨 A20-Luc-GFP 세포주를 96 웰 플레이트에 200μl 용량으로 1Х10
2세포수부터 1x10
5세포수까지 분주하고 루시페린을 150ug/ml농도로 주입한 뒤 생물학적발광을 측정하여 세포수에 따른 생물학적발광 정도를 확인하였다. 종양 이식 마우스에 루시페린을 150mg/kg로 복강내 주사하고 루시퍼라제를 발현하는 종양의 생물학적발광을 측정하여 생체 내 종양 성장 정도를 확인하였다. 생물학적발광은 생체 내 형광분광 분석기(IVIS Lumina XRMS)를 통해 측정하였다.The A20-Luc-GFP cell line suspended in RPMI medium was dispensed in a 96-well plate at a volume of 200 μl from 1Х10 2 cells to 1x10 5 cells, luciferin was injected at a concentration of 150ug/ml, and bioluminescence was measured to determine the The degree of bioluminescence was confirmed. Luciferin was intraperitoneally injected into tumor-implanted mice at 150 mg/kg, and the bioluminescence of the luciferase-expressing tumor was measured to determine the degree of tumor growth in vivo. Bioluminescence was measured using an in vivo fluorescence spectrometer (IVIS Lumina XRMS).
본 발명의 TLR5 작용제에 의한 단독 또는 병용 투여에 따른 항암 효과를 관찰하기 위하여. 종양 마우스 모델을 유도한 후 투여 물질 별 종양의 성장을 비교하였다.To observe the anticancer effect of the TLR5 agonist of the present invention administered alone or in combination. After induction of the tumor mouse model, the growth of the tumor for each administered material was compared.
실험 결과Experiment result
1. 대장암 마우스 모델 1. Colorectal Cancer Mouse Model
(1) 종양 마우스 모델에서 TLR5 작용제에 의한 항암 효과 검증(1) Verification of anticancer effect by TLR5 agonist in tumor mouse model
본 발명의 TLR5 작용제에 의한 단독 또는 병용 투여에 따른 항암 효과를 관찰하기 위하여. 종양 마우스 모델을 유도한 후 투여 물질 별 종양의 성장을 비교하였다.To observe the anticancer effect of the TLR5 agonist of the present invention administered alone or in combination. After induction of the tumor mouse model, the growth of the tumor for each administered material was compared.
먼저, MC-38 세포주가 이식된 마우스의 복강 내에 이식 후 6일째부터 3일 간격으로 총 3회씩 생리식염수 200 μl/kg (대조군: ●), TLR5 작용제 100 μg/kg (α, 항 PD-1 200 μg/mice (▲)를 각 단독으로 투여하고 항 PD-1 및 TLR5 작용제 (▼)를 병용 투여하여 항암효과를 관찰하였고, 상기 결과를 도 2에 나타내었다. 도 2에 나타낸 바와 같이, 본원발명의 TLR5 작용제는 종양의 성장 속도를 지연시켰으며 이 같은 항암효과는 양성대조군인 항 PD-1 보다 종양 생장이 현저하게 억제된 것을 확인할 수 있었다. 또한, 상기 실험 결과를 통하여, 본 발명의 TLR5 작용제 및 항 PD-1의 병용 투여는 각 물질의 단독 투여와 대비하여 항종양에 상승된 시너지 효과가 있음을 알 수 있다.First, 200 μl/kg of physiological saline (control group: ●), 100 μg/kg of TLR5 agonist (α, anti-PD-1 200 μg/mice (▲) was administered alone and anti-PD-1 and TLR5 agonists (▼) were administered in combination to observe the anticancer effect, and the results are shown in Figure 2. As shown in Figure 2, the present application The TLR5 agonist of the present invention delayed the growth rate of the tumor, and it was confirmed that this anticancer effect significantly inhibited the tumor growth than the positive control, anti-PD-1. It can be seen that the combined administration of the agonist and the anti-PD-1 has an increased synergistic effect on the antitumor compared to the administration of each substance alone.
한편, MC-38 세포주가 복강 내에 이식된 마우스를 모니터링하였고, 그 결과를 도 3에 나타내었다. 마우스에서의 암의 크기는 음성 또는 양성대조군과 비교하였을 때, 본원발명의 TLR5 작용제 투여로 인하여 작아진 것이 관찰 가능함을 알 수 있다. 이러한 결과는 본 발명의 TLR5 작용제("KMRC011")가 면역관문억제제로서 사용되어 효과적인 항암제로 사용 가능할 뿐만 아니라, 기존 면역관문억제제와 병용 투여되어 종양 치료에 시너지 효과를 가지는 항암보조제로서 사용될 수 있음을 의미한다.Meanwhile, mice transplanted with the MC-38 cell line intraperitoneally were monitored, and the results are shown in FIG. 3 . It can be seen that it can be observed that the size of the cancer in mice is reduced due to administration of the TLR5 agonist of the present invention when compared to the negative or positive control group. These results indicate that the TLR5 agonist ("KMRC011") of the present invention can be used as an immune checkpoint inhibitor and can be used as an effective anticancer agent, and can be used as an anticancer adjuvant having a synergistic effect in tumor treatment when administered in combination with an existing immune checkpoint inhibitor. it means.
(2) TLR5 작용제에 의한 종양 미세환경 조절 확인(2) Confirmation of tumor microenvironment regulation by TLR5 agonists
본 발명의 TLR5 작용제가 면역관문억제제로서 종양 미세환경 변화를 유도시키는지를 확인하기 위해, 종양 마우스 모델에서 투여 물질 별 마우스 비장과 림프절 세포의 면역 프로파일링을 유세포 분석으로 평가하고자 하였다. 이를 위하여 본 발명자들은 마우스 비장과 림프절 세포로부터 M1 및 M2 대식세포의 분화 조건을 확립하였다. 일반적으로 M1 대식세포는 종양 공격성을 나타내어 항암 효과가 있는 것으로 알려졌고, M2 대식세포는 암에 친화적인 종양 지지형 대식세포로서 종양을 성장시키는 것으로 알려져 있다. 따라서 상기 M1/M2 대식세포의 분극도 변화를 통하여 본 발명의 TLR5 작용제가 종양 세포주의 종양 미세환경을 조절하여 항암 효과가 있는지를 확인하였다.In order to confirm whether the TLR5 agonist of the present invention induces changes in the tumor microenvironment as an immune checkpoint inhibitor, the immune profiling of mouse spleen and lymph node cells for each administered substance in a tumor mouse model was evaluated by flow cytometry. To this end, the present inventors established conditions for differentiation of M1 and M2 macrophages from mouse spleen and lymph node cells. In general, M1 macrophages are known to have an anticancer effect by showing tumor aggression, and M2 macrophages are known to grow tumors as tumor-supporting macrophages that are friendly to cancer. Therefore, it was confirmed whether the TLR5 agonist of the present invention had an anticancer effect by regulating the tumor microenvironment of the tumor cell line through the change in the degree of polarization of the M1/M2 macrophages.
종양 동물 모델에서 생리식염수, TLR5 작용제 및 항 PD-1을 각 단독으로 투여, 항 PD-1 및 TLR5 작용제를 병용하여 투여한 후 마우스 비장과 림프절 세포로부터 M1 및 M2 대식세포를 유세포 분석기를 이용하여 관찰하여 분극도(polarization)를 확인하였고, 그 결과를 도 4 및 도 5에 나타내었다. TLR5 작용제를 처리한 비장과 림프절 세포의 경우, M1 대식세포 (F4/80+ CD206-)의 분극도는 증가시키는 반면 M2 대식세포(F4/80+ CD206+)의 분극도는 감소시키는 것으로 확인되었다. 이처럼 TLR5 작용제는 면역관문억제제의 반응을 높이기 위하여 면역세포아형인 M1 및 M2 대식세포의 분포를 조절할 수 있음을 확인할 수 있다.In a tumor animal model, after administration of saline, a TLR5 agonist and anti-PD-1 alone, and an anti-PD-1 and TLR5 agonist in combination, M1 and M2 macrophages from mouse spleen and lymph node cells were analyzed using flow cytometry. The polarization was confirmed by observation, and the results are shown in FIGS. 4 and 5 . In the case of spleen and lymph node cells treated with TLR5 agonist, it was confirmed that the polarization of M1 macrophages (F4/80+ CD206-) was increased, while that of M2 macrophages (F4/80+ CD206+) was decreased. As such, it can be confirmed that the TLR5 agonist can regulate the distribution of immune cell subtypes M1 and M2 macrophages in order to increase the response of the immune checkpoint inhibitor.
따라서, 본 발명의 TLR5 작용제("KMRC011")는 단독 또는 기존의 면역관문억제제와 병용하여 종양 미세환경을 변화시킴으로서 항종양 효과를 최적으로 발휘할 수 있는 환경을 만들 수 있음을 알 수 있다.Therefore, it can be seen that the TLR5 agonist ("KMRC011") of the present invention can create an environment in which the antitumor effect can be optimally exerted by changing the tumor microenvironment either alone or in combination with an existing immune checkpoint inhibitor.
2. B 세포 림프종 마우스 모델2. B-Cell Lymphoma Mouse Model
(1) A20-Luc-GFP 세포주의 생물학적발광 검증(1) Verification of bioluminescence of A20-Luc-GFP cell line
A20-Luc-GFP 세포주를 1Х10
2세포수부터 1x10
5세포수까지 각기 다른 세포수의 생물학적발광을 측정하여 세포수에 따른 생물학적발광 정도를 확인하였고 그 결과를 도 7에 나타내었으며, 세포수가 증가함에 따라 생물학적발광 정도도 비례하여 증가함이 확인되었다. 이러한 결과는 A20-Luc-GFP 세포주를 사용하여 종양 마우스 모델을 도입하였을 때 형성된 종양의 생물학적발광을 측정할 경우, 생체 내 종양 성장 정도를 확인할 수 있음을 의미한다.In the A20-Luc-GFP cell line, the degree of bioluminescence according to the number of cells was confirmed by measuring the bioluminescence of different cell numbers from 1Х10 2 cells to 1x10 5 cells. Accordingly, it was confirmed that the degree of bioluminescence increased in proportion. These results mean that when the bioluminescence of the tumor formed when the tumor mouse model is introduced using the A20-Luc-GFP cell line is measured, the degree of tumor growth in vivo can be confirmed.
(2) 종양 마우스 모델에서 TLR5 작용제에 의한 항암 효과 검증(2) Verification of anticancer effect by TLR5 agonist in tumor mouse model
본 발명의 TLR5 작용제("KMRC011")의 단독 또는 병용 투여에 따른 항암 효과를 관찰하기 위하여, 종양 마우스 모델을 유도한 후 투여 물질 별 종양의 성장을 비교하였다.In order to observe the anticancer effect of the TLR5 agonist of the present invention ("KMRC011") alone or in combination, the tumor growth was compared for each administered substance after inducing a tumor mouse model.
마우스의 피하에 A20-Luc-GFP 세포주의 이식 후 9일째부터 3일 간격으로 총 3회씩 생리식염수 200 μl/mice, TLR5 작용제 100 μg/kg, 항 PD-1 200 μg/mice를 각 단독으로 투여하고 항 TLR5 작용제 및 항 PD-1을 병용 투여하였다. 생물학적발광을 측정하여 항암효과를 관찰하였고, 상기 결과를 도 6에 나타내었다. 도 8 내지 9에 나타낸 바와 같이, 본원발명의 TLR5 작용제는 종양의 성장 속도를 지연시켰다. 또한, 상기 실험 결과를 통하여, 본 발명의 TLR5 작용제 및 항 PD-1의 병용 투여는 각 물질의 단독 투여와 대비하여 항종양에 상승된 시너지 효과가 있음을 알 수 있다.After subcutaneously transplanting the A20-Luc-GFP cell line into the mouse, 200 μl/mice of physiological saline, 100 μg/kg of TLR5 agonist, and 200 μg/mice of anti-PD-1 were administered alone 3 times at intervals of 3 days from the 9th day. and anti-TLR5 agonist and anti-PD-1 were administered in combination. The anticancer effect was observed by measuring bioluminescence, and the results are shown in FIG. 6 . 8 to 9 , the TLR5 agonist of the present invention retarded the growth rate of tumors. In addition, through the above experimental results, it can be seen that the combined administration of the TLR5 agonist of the present invention and the anti-PD-1 has an increased synergistic effect on the antitumor compared to the administration of each substance alone.
A20-Luc-GFP 세포주가 이식된 마우스의 형성된 종양의 크기를 BLI로 측정하였고, 그 결과를 도 9에 나타냈다. 마우스에서의 암의 크기는 음성대조군(●) TLR5 작용제 투여군(■), 항 PD-1 투여군(▲)과 비교하였을 때, 본 발명의 TLR5 작용제 및 항 PD-1의 병용 투여(▼)로 인하여 작아진 것이 관찰 가능함을 알 수 있다.The size of the tumor formed in mice transplanted with the A20-Luc-GFP cell line was measured by BLI, and the results are shown in FIG. 9 . Compared to the negative control group (●), the TLR5 agonist administration group (■), and the anti-PD-1 administration group (▲), the size of the cancer in mice was due to the combined administration (▼) of the TLR5 agonist and anti-PD-1 of the present invention. It can be seen that the smaller ones are observable.
이러한 결과는 본 발명의 TLR5 작용제("KMRC011")가 단독 항종양효과를 보일 뿐만 아니라, 기존 면역관문억제제와 병용 투여시 종양 치료에 시너지 효과를 가지는 항암보조제로서 사용될 수 있음을 규명하였다.These results confirmed that the TLR5 agonist ("KMRC011") of the present invention not only exhibits a single antitumor effect, but can also be used as an anticancer adjuvant having a synergistic effect in tumor treatment when administered in combination with an existing immune checkpoint inhibitor.
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.As described above in detail a specific part of the present invention, for those of ordinary skill in the art, it is clear that this specific description is only a preferred embodiment, and the scope of the present invention is not limited thereby. something to do. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.
본 발명은 플라젤린으로부터 유도된 TLR5 작용제를 유효성분으로 포함하는 항암 제 조성물에 관한 것으로, 본 발명의 플라젤린으로부터 유도된 TLR5 작용제는 단독 또는 면역관문억제제와 동시에 항암 또는 항암 보조의 효과를 나타낸다.The present invention relates to an anticancer composition comprising a flagellin-derived TLR5 agonist as an active ingredient, wherein the flagellin-derived TLR5 agonist of the present invention exhibits an anticancer or anticancer adjuvant effect either alone or at the same time as an immune checkpoint inhibitor.
Claims (12)
- 플라젤린으로부터 유도된 TLR5 작용제의 치료학적 유효량을 포함하는 항암제 조성물.An anticancer composition comprising a therapeutically effective amount of a TLR5 agonist derived from flagellin.
- 제 1 항에 있어서, 상기 플라젤린으로부터 유도된 TLR5 작용제는 플라젤린의 D0 도메인 및 D1 도메인을 포함하는 것을 특징으로 하는 조성물.The composition of claim 1 , wherein the TLR5 agonist derived from flagellin comprises a D0 domain and a D1 domain of flagellin.
- 제 1 항에 있어서, 상기 플라젤린으로부터 유도된 TLR5 작용제는 D1 도메인 내부에 서열번호 1에 개시된 아미노산 서열의 링커 펩타이드를 포함하는 것을 특징으로 하는 조성물.The composition according to claim 1, wherein the flagellin-derived TLR5 agonist comprises a linker peptide of the amino acid sequence set forth in SEQ ID NO: 1 inside the D1 domain.
- 제 1 항에 있어서, 상기 플라젤린으로부터 유도된 TLR5 작용제는 서열번호 2에 개시된 아미노산 서열을 포함하는 것을 특징으로 하는 조성물.The composition of claim 1, wherein the flagellin-derived TLR5 agonist comprises the amino acid sequence set forth in SEQ ID NO:2.
- 제 1 항에 있어서, 상기 플라젤린으로부터 유도된 TLR5 작용제는 종양 미세환경을 조절하는 것을 특징으로 하는 조성물.The composition of claim 1, wherein the flagellin-derived TLR5 agonist modulates the tumor microenvironment.
- 제 1 항에 있어서, 상기 플라젤린으로부터 유도된 TLR5 작용제는 비장과 림프절에서 M1 (F4/80+CD206-) 분극을 증가시키고 M2 (F4/80+CD206+) 분극을 감소시키는 것을 특징으로 하는 조성물.The composition of claim 1 , wherein the flagellin-derived TLR5 agonist increases M1 (F4/80+CD206−) polarization and decreases M2 (F4/80+CD206+) polarization in the spleen and lymph nodes.
- 제 1 항에 있어서, 상기 조성물은 암세포의 성장 저해 활성을 갖는 것을 특징으로 하는 조성물.The composition according to claim 1, wherein the composition has a growth inhibitory activity of cancer cells.
- 제 1 항에 있어서, 면역관문억제제를 추가로 포함하는 것을 특징으로 하는 조성물.The composition according to claim 1, further comprising an immune checkpoint inhibitor.
- 제 8 항에 있어서, 상기 면역관문억제제는 항 PD-1 항체인 것을 특징으로 하는 조성물.The composition of claim 8, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody.
- 제 9 항에 있어서, 상기 항 PD-1 항체는 아벨루맙(avelumab), 트리멜리무맙(Tremelimumab), 이필리무맙(Ipilimumab), 니볼루맙(Nivolumab), 펨브롤리주맙(Pembrolizumav), 아테졸리주맙(Atezolizumab), 두발루맙(Durvalumab), 람브로리주맙(Lamvrolizumab), AMP-224, MEDI4376 및 CT-011로 이루어진 군으로부터 선택되는 1종 이상인 것을 특징으로 하는 조성물.The method of claim 9, wherein the anti-PD-1 antibody is avelumab, trimelimumab, ipilimumab, nivolumab, pembrolizumab, atezolizumab ( Atezolizumab), duvalumab (Durvalumab), lambrolizumab (Lamvrolizumab), AMP-224, MEDI4376 and a composition characterized in that at least one selected from the group consisting of CT-011.
- 제 1 항 내지 제 10 항 중 어느 한 항에 있어서, 약제학적으로 허용되는 담체를 포함하는 것을 특징으로 하는 조성물.11. The composition according to any one of claims 1 to 10, comprising a pharmaceutically acceptable carrier.
- 제 11 항의 조성물을 포함하는 약학적 제제.A pharmaceutical formulation comprising the composition of claim 11 .
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20120104177A (en) * | 2009-10-06 | 2012-09-20 | 파나셀라 랩스, 아이엔씨. | Use of toll-like receptor and agonist for treating cancer |
WO2016146261A1 (en) * | 2015-03-16 | 2016-09-22 | Amal Therapeutics Sa | Combination of an immune checkpoint modulator and a complex comprising a cell penetrating peptide, a cargo and a tlr peptide agonist for use in medicine |
KR101744297B1 (en) * | 2016-06-21 | 2017-06-07 | 한국생산기술연구원 | Method for production of TLR5 agonist |
US10202426B2 (en) * | 2014-07-30 | 2019-02-12 | Genome Protection, Inc. | Flagellin compositions and uses |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
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KR102024322B1 (en) | 2018-02-22 | 2019-09-23 | 경희대학교 산학협력단 | A composition for enhancing radiation sensitivity and an anticancer adjuvent composition comprising ginsenoside rg3 |
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Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20120104177A (en) * | 2009-10-06 | 2012-09-20 | 파나셀라 랩스, 아이엔씨. | Use of toll-like receptor and agonist for treating cancer |
US10202426B2 (en) * | 2014-07-30 | 2019-02-12 | Genome Protection, Inc. | Flagellin compositions and uses |
WO2016146261A1 (en) * | 2015-03-16 | 2016-09-22 | Amal Therapeutics Sa | Combination of an immune checkpoint modulator and a complex comprising a cell penetrating peptide, a cargo and a tlr peptide agonist for use in medicine |
KR101744297B1 (en) * | 2016-06-21 | 2017-06-07 | 한국생산기술연구원 | Method for production of TLR5 agonist |
Non-Patent Citations (2)
Title |
---|
GONZALEZ CALEB, WILLIAMSON SARAH, GAMMON SETH, PIWNICA-WORMS DAVID: "TLR5 agonists enhance anti-tumor immunity and overcome resistance to immune checkpoint therapy", CANCER RESEARCH, 1 April 2020 (2020-04-01), XP055876771, DOI: 10.1158/1538-7445.AM2020-921Published * |
NICHOLAS D. LEIGH, GUANGLIN BIAN, XILAI DING, HONG LIU, SEMRA AYGUN-SUNAR, LYUDMILA G. BURDELYA, ANDREI V. GUDKOV, XUEFANG CAO: "A Flagellin-Derived Toll-Like Receptor 5 Agonist Stimulates Cytotoxic Lymphocyte-Mediated Tumor Immunity", PLOS ONE, vol. 9, no. 1, pages e85587, XP055391192, DOI: 10.1371/journal.pone.0085587 * |
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JP2023504907A (en) | 2023-02-07 |
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